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direct / indirect detection in WB - (Oct/04/2006 )

Hi,

For some purposes, sometimes I do a fluorescence WB with a secondary Ab labelled with FITC. But what are the advantages / disadvantages (if they exist) of doing this versus using the primary Ab directly labelled with FITC? unsure.gif

thanks a lot rolleyes.gif

-pumuki-

you amplify the signal due to multiplicity of binding sites for the 2nd antibody on the 1st antibody.

-mdfenko-

mdfenko is right, moreover it is known that secondary and third antibodies (as in ELISA) rise specificity of signal (better signal/noise ratio)

-The Bearer-

Thanks a lot guys, smile.gif smile.gif smile.gif
now I think I´ve heard that explanation before when I was at University and somebody told us something like that.

-pumuki-